• 제목/요약/키워드: Glutathione dependent enzyme

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Alterations of Glutathione and Glutathione-Dependent Enzyme Activities by Monosodium-L-Glutamate in Rats with Carbon Tetrachloride-Induced Liver Damage (사염화탄소와 Monosodium-L-Glutamate 병용투여에 의한 간조직의 환원형글루타치온 함량 및 그의 관련효소활성의 변화)

  • 김형춘;이왕섭;전완주;김수희;주왕기
    • YAKHAK HOEJI
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    • v.35 no.5
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    • pp.384-388
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    • 1991
  • To explore the effect of monosodium-L-glutamate(MSG) on CCI$_{4}$-damaged liver in Wister male rat, 5% MSG solution as drink water were administered after S.C. injection of 0.1 mg/kg CC1$_{4}$ twice a week for 4 weeks. After last administration of MSG, heptic glutathione(GSH) dependent system was assayed. It showed that MSG increased significanly hepatic glutathione(GSH) and glutathione peroxidase(GSH$_{px}$), but decreased glutathione-S-transferase(GST) acivity in normal rats. MSG increased significantly the GSH$_{px}$ and GST activities in rats with CCI$_{4}$-induced liver damage. These results indicate that decrease of GSH dependent systems in CC1$_{4}$ liver injury might be partially elevated by coadministration of MSG.

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Analysis and Characterization of Glutathione Peroxidases in an Environmental Microbiome and Isolated Bacterial Microorganisms

  • Yun-Juan Bao;Qi Zhou;Xuejing Yu;Xiaolan Yu;Francis J. Castellino
    • Journal of Microbiology and Biotechnology
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    • v.33 no.3
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    • pp.299-309
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    • 2023
  • Glutathione peroxidases (Gpx) are a group of antioxidant enzymes that protect cells or tissues against damage from reactive oxygen species (ROS). The Gpx proteins identified in mammals exhibit high catalytic activity toward glutathione (GSH). In contrast, a variety of non-mammalian Gpx proteins from diverse organisms, including fungi, plants, insects, and rodent parasites, show specificity for thioredoxin (TRX) rather than GSH and are designated as TRX-dependent peroxiredoxins. However, the study of the properties of Gpx in the environmental microbiome or isolated bacteria is limited. In this study, we analyzed the Gpx sequences, identified the characteristics of sequences and structures, and found that the environmental microbiome Gpx proteins should be classified as TRX-dependent, Gpx-like peroxiredoxins. This classification is based on the following three items of evidence: i) the conservation of the peroxidatic Cys residue; ii) the existence and conservation of the resolving Cys residue that forms the disulfide bond with the peroxidatic cysteine; and iii) the absence of dimeric and tetrameric interface domains. The conservation/divergence pattern of all known bacterial Gpx-like proteins in public databases shows that they share common characteristics with that from the environmental microbiome and are also TRX-dependent. Moreover, phylogenetic analysis shows that the bacterial Gpx-like proteins exhibit a star-like radiating phylogenetic structure forming a highly diverse genetic pool of TRX-dependent, Gpx-like peroxidases.

Overexpression, Purification, and Biochemical Characterization of the Thermostable NAD-dependent Alcohol Dehydrogenase from Bacillus stearothermophilus

  • Shim, Eun-Jung;Jeon, Sang-Hoon;Kong, Kwang-Hoon
    • Journal of Microbiology and Biotechnology
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    • v.13 no.5
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    • pp.738-744
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    • 2003
  • The gene ADH encoding NAD-dependent alcohol dehydrogenase from Bacillus stearothennophilus was cloned and overexpressed as a GST fusion protein at a high level in Escherichia coli. The expressed fusion protein was purified simply by glutathione affinity chromatography. GST fusion protein was then cleaved by thrombin, while soluble enzyme was further purified by glutathione affinity chromatography. The recombinant enzyme had the same elctrophoretic mobility as the native enzyme from Bacillus stearothennophilus. The recombinant enzyme catalyzed the oxidation of a number of alcohols and exhibited high activities towards secondary alcohols. The $K_m\;and\;V_{max}$ values of the recombinant enzyme for ethanol were 5.11 mM and 61.35 U/mg, respectively. Pyridine and imidazole notably inhibited the enzymatic activity. The activity of the recombinant enzyme optimally proceeded at pH 9.0 and $70^{\circ}C$. The midpoint of the temperature-stability curve for the recombinant enzyme was approximately $68^{\circ}C$, and the enzyme was not completely inactivated even at $85^{\circ}C$. The recombinant enzyme showed a high resistance towards denaturing agents (0.05% SDS, 0.1 M urea). Therefore, due to its stability and relatively broad substrate specificity, the recombinant enzyme could be utilized in bio-industrial processes and biosensors.

Protective Effects of Black Rice Extracts on Oxidative Stress Induced by tert-Butyl Hydroperoxide in HepG2 Cells

  • Lee, Seon-Mi;Choi, Youngmin;Sung, Jeehye;Kim, Younghwa;Jeong, Heon-Sang;Lee, Junsoo
    • Preventive Nutrition and Food Science
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    • v.19 no.4
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    • pp.348-352
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    • 2014
  • Black rice contains many biologically active compounds. The aim of this study was to investigate the protective effects of black rice extracts (whole grain extract, WGE and rice bran extract, RBE) on tert-butyl hydroperoxide (TBHP)-induced oxidative injury in HepG2 cells. Cellular reactive oxygen species (ROS), antioxidant enzyme activities, malondialdehyde (MDA) and glutathione (GSH) concentrations were evaluated as biomarkers of cellular oxidative status. Cells pretreated with 50 and $100{\mu}g/mL$ of WGE or RBE were more resistant to oxidative stress in a dose-dependent manner. The highest WGE and BRE concentrations enhanced GSH concentrations and modulated antioxidant enzyme activities (glutathione reductase, glutathione-S-transferase, catalase, and superoxide dismutase) compared to TBHP-treated cells. Cells treated with RBE showed higher protective effect compared to cells treated with WGE against oxidative insult. Black rice extracts attenuated oxidative insult by inhibiting cellular ROS and MDA increase and by modulating antioxidant enzyme activities in HepG2 cells.

Increased antioxidant enzyme activities and scavenging effects of oxygen free radicals by Cheongahwan (청아환(靑娥丸)에 의한 활성(活性) 산소류(酸素類)의 소거(消去) 작용(作用)과 항산화(抗酸化) 효소계(酵素系)의 활성(活性) 증가(增加) 효과(效果)에 대(對)한 연구(硏究))

  • Jeong, Ji-Cheon
    • The Journal of Korean Medicine
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    • v.18 no.2
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    • pp.355-365
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    • 1997
  • This study was undertaken to examine the effect of Cheongahwan(CAH), being known to reinforce Kidney-yang, on the activities of endogenous antioxidant enzymes and the production of oxygen free radicals in the kidney tissues. Alterations in enzyme activities were observed after in vivo treatment in rats. CAH caused a significant increase in the activities of superoxide dismutase (SOD), glutathione peroxidase and glutathione S-transferase. But catalase activity was not significantly altered by CAH. Treatment in vitro of CAH decreased the production of oxygen free radicals in a dose-dependent fashion. These results suggest that CAH stimulate the activities of antioxidant enzymes and inhibit directly the production of oxygen free radicals. These effects of CAH may contribute to prevent the oxygen free radical-induced impairment of cell function.

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Change of ROS Generation and Antioxidant Enzyme Activity of Flavonol Quercetin in the Presence of Vitamin E, L-Ascorbit acid, Reduced Glutathione on the B16F10 Murine Melanoma Cells (B16F10 세포에서 Quercetin과 Vitamin E, L-Ascorbic acid, 환원형 글루타치온과의 병용 투여에 의한 활성산소종 발생과 항산화 효소의 활성 변화)

  • 허정심;김안근
    • YAKHAK HOEJI
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    • v.47 no.6
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    • pp.432-437
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    • 2003
  • It has been known that quercetin, a bioflavonoid widely distributed in fruits and vegetables as dietary-derived flavonoid and exert significant multiple biological effects such as antioxidant and anti-inflammatory, anti-tumor effects. In addition, it has been shown to have a chemoprotective role in cancer, though complex effects on signal transduction involved in cell proliferation and angiogenesis. The present study investigated whether quercetin can enhance antioxidant enzyme activity (glutathione peroxidase: GPx, superoxide dismutase: SOD, catalase: CAT) and regulate the reactive oxygen species (ROS) generation in the presence of vitamin E, L-ascorbic acid, reduced glutathione (GSH) on B16F10 murine melanoma cells. After 48h treatment of cells with quercetin in the presence of vitamin E, L-ascorbic acid, GSH, we measured the cytotoxicities by MTT assay. The cells exhibited a dose-dependent inhibition in their proliferation in the presence of vitamin E, L-ascorbic acid, GSH respectively. We also investigated the effects of antioxidant enzyme activity and ROS generation. The antioxidant enzyme activity of quercetin in the presence of vitamin E was stronger than GSH, L-ascorbic acid, the same treatments decreased ROS generation in B16F10 murine melanoma cells. Taken together, these result demonstrate that the antioxidant effect of quercetin can enhanced in the presence of vitamin E and it might plays an important role in anti-oxidative effects.

Effect of Prunella vulgaris L. on Chemopreventive Enzymes of Colorectal Cancer (꿀풀하고초가 직장암 예방효소 활성에 미치는 영향)

  • Shon, Yun-Hee;Seo, Jae-Beom;Nam, Kyung-Soo
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.22 no.1
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    • pp.126-130
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    • 2008
  • Water extract from Prunella vulgaris L. (PVW) was tested for colon cancer chemopreventive activity by measuring the activities of cytochrome P450 1A1, phase Ⅱ detoxification enzyme [quinone reductase (QR) and glutathione S-transferase (GST)] and ornithine decarboxylase (ODC) and glutathione (GSH) levels in cultured human colorectal adenocarcinoma HT-29 cells. PVW significantly inhibited 7,12-dimethylbenz[a]anthracene (DMBA)-induced cytochrome P450 1A1 activity at 10 and 50 ${\mu}g/ml$. PVW induced QR activity in a dose-dependent manner over a concentration range of $1{\sim}50\;{\mu}g/ml$. GST activity was also induced with the treatment of PVW in HT-29 cells. In addition GSH levels were increased with PVW. PVW inhibited ODC activity, a key enzyme of polyamine biosynthesis, which is enhanced in tumor promotion. These results suggest that Prunella vulgaris L. has colon cancer chemopreventive activity by inhibiting cytochrome P450 1A1 and ODC activities and by increasing phase Ⅱ enzyme activity and GSH levels.

Reciprocal Effect of DHEA and Rietary Fat on Glutathione Utilizing Detoxifying System in Rat Liver Tissue

  • Kwak, Chung-Shil;Kwon, In-Soon;Park, Sang-Chul
    • Nutritional Sciences
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    • v.3 no.1
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    • pp.11-17
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    • 2000
  • This study was intended to examine whether dehydroepiandrosterone (DHEA) and dietary fat level or source could modulate glutathione utilizing detoxifying system activity and the cytosolic NADPH generation in rat liver. Male Sprague-Dawley rats were fed semipurifed diet containing either 2%(w/w) corn oil (low level of corn oil diet: 5 ca% of fat) 15% corn oil (high level of corn oil diet: 31 cal% of fat) or 13% sardine oil plus 2% corn oil(high level of fish oil diet: 31 cal% of fat) for 9 weeks. Half of the rats in each diet group were fed a diet supplemented with 0.2% DHEA (w/w). DHEA administration increased plasma total cholesterol level in low corn oil diet-fed rats. The high fish oil diet significantly decreased plasma total cholesterol level compared to the high corn oil diet. Plasma triglyceride level was not significantly changed by DHEA administration and dietary fat level and source. Fasting plasma glucose level was increased by DHEA administration and fish oil diet. Glucose 6-phosphate dehydrogenase activity in liver tissue was significantly increased by DHEA administration and high fat diet, especially fish oil diet. Malic enzyme activity in liver tissue was significantly increased by DHEA administration and high fat diet, especially fish oil diet. Malic enzyme activity in liver tissue was significantly increased by DHEA administration. DHEA suppressed the glutathione peroxidase, glutathione-dependent enzymes compared to the low corn oil diet, while fish oil diet elevated the activity of glutathione peroxidase and glutathione reductase compared to corn oil diet. These results suggest that DHEA administration and high level of corn oil diet may suppress the cellular detoxifying system activity through reduction of glutathione utilization, while the fish oil diet did not show these effects.

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Effect of Cnidii Rhizoma on Phase II Enzyme and Ornithine Decarboxylase Activities (천궁이 Phase II 효소 유도와 Ornithine Decarboxylase 활성에 미치는 영향)

  • Shon, Yun-Hee;Kim, Mee-Kyung;Cho, Hyun-Jung;Nam, Kyung-Soo
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.20 no.6
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    • pp.1572-1575
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    • 2006
  • Water extract from Cnidii Rhizoma (CRW) was tested for colon cancer chemopreventive activity by measuring the induction of phase II detoxification enzyme activity [quinone reductase (QR) and glutathione S-transferase (GST)] and glutathion (GSH) levels and ornithine decarboxylase (ODC) activity in cultured human colorectal adenocarcinoma HT-29 cells. CRW inhibited cell proliferation in cultured HT-29 cells. CRW induced QR activity in a dose-dependent manner in a concentration range of 0.1${\sim}$5.0 $mg/m{\ell}$. GST activity was also induced with the treatment of CRW in HT-29 cells. In addition GSH levels was increased with CRW. CRW inhibited ODC activity, a key enzyme of polyamine biosynthesis, which is enhanced in tumor promotion. These results suggest that CRW has colon cancer chemopreventive activity by increasing phase II enzyme activity and GSH levels and inhibiting ODC activity in vitro.

Effect of Se-methylselenocysteine on the Antioxidant System in Rat Tissues

  • Shin, Ho-Sang;Choi, Eun-Mi
    • Preventive Nutrition and Food Science
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    • v.15 no.4
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    • pp.267-274
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    • 2010
  • We assessed the effect of Se-methylselenocysteine (MSC) treatment, at a dose of 0.75 mg/rat/day for 1 or 2 weeks, on the activities of antioxidant systems in Sprague-Dawley rat tissues. Significant changes in glutathione and antioxidant enzyme activities, with different patterns among tissues, were evidenced. Glutathione content and its reduction state in the liver, lung, and kidney were elevated upon MSC treatment, whereas they were significantly lowered in the spleen. Among the tissues exhibiting glutathione increase, there were different enzymatic responses: $\gamma$-glutamylcysteine ligase activity, the rate-limiting enzyme in the glutathione synthesis pathway, was increased in the liver, whereas the activities of the enzymes associated with glutathione recycling, namely, glutathione peroxidase, glutathione reductase, and glucose 6-phosphate dehydrogenase, were significantly increased in the lung and the kidney. The superoxide dismutase activity was decreased in all tissues upon MSC treatment, whereas catalase activity was increased in all tissues but the liver. Lipid peroxidation level was transiently increased at 1 week in the lung and the kidney, whereas it was persistently increased in the spleen. The increase was not evident in the liver. The results indicate that the MSC treatment results in an increase in the antioxidant capacity of the liver, lung, and kidney principally via an increase in glutathione content and reduction, which appeared to be a result of increased synthesis or recycling of glutathione via tissue-dependent adaptive response to oxidative stress triggered by MSC. The spleen appeared to be very sensitive to oxidative stress, and therefore, the adaptive response could not provide protection against oxidative damage.